Journal: bioRxiv

Article Title: Aberrant cortical development is driven by impaired cell cycle and translational control in a DDX3X syndrome model

doi: 10.1101/2022.02.21.481343

Figure Lengend Snippet: ( A ) RT-qPCR quantification of Ddx3x and Ddx3y mRNA levels in FACS-isolated Tdtomato+ cells from cKO male E11.5 cortices. ( B ) Schematic of Ddx3y CRISPR sgRNA electroporation of E14.5 brain. ( C ) RT-qPCR quantification of Ddx3y and Ddx3x levels in GFP+ FACS-isolated cells from E17.5 male and female mice electroporated with pCAG-GFP and either no sgRNA or Ddx3y sgRNA. ( D ) Representative sections of E17.5 male brains electroporated at E14.5 with pCAG-GFP and either no sgRNA or Ddx3y sgRNA and stained with anti-GFP (grey). Dotted lines, ventricular and pial surfaces; brackets delineate equivalently sized bins. ( E ) Quantification of distribution of GFP+ cells. ( F ) Same as ( D ), but sections were stained with anti-GFP (grey), Sox2 (magenta), and Neurod2 (red). ( G, H ) Quantification of GFP co-localization with Sox2 ( G ) or Neurod2 ( H ). Scale bars, indicated. Error bars, S.D. **p<0.01, ***p<0.001.

Article Snippet: The following primary antibodies were used, rat: anti-SOX2 (Thermo Fisher, 14-9811-82, 1:1000), anti-BrdU (Abcam, ab6326, 1:200); mouse: anti-TUJ1 (Biolegend, 801202, 1:2000), anti-CTIP2 (Abcam, c8035, 1:500); rabbit: anti-TBR2 (Abcam, AB23345, 1:1000), anti-CC3 (Cell Signaling, 9661, 1:250), anti-NEUROD2 (Abcam, AB104430, 1:500), anti-Ki67 (Cell Signaling Technology, 12202, 1:1000), anti-PH3 (Millipore, 06-570, 1:500), anti-TBR1 (Cell Signaling Technology, 49661S, 1:1000), anti-Lhx2 (Millipore, ABE1402, 1:500), anti-DDX3X (Sigma Aldrich, HPA001648, 1:500); chicken anti-GFP (Abcam, Ab13970, 1:1000).

Techniques: Quantitative RT-PCR, Isolation, CRISPR, Electroporation, Staining

Journal: bioRxiv

Article Title: Aberrant cortical development is driven by impaired cell cycle and translational control in a DDX3X syndrome model

doi: 10.1101/2022.02.21.481343

Figure Lengend Snippet: ( A ) RT- qPCR quantification of Setd3 knockdown in N2A cells. ( B ) Representative sections of E17.5 brains from mice electroporated at E14.5 with pCAG-GFP and scrambled or Setd3 siRNAs and immunostained with GFP (grey), Sox2 (magenta), Neurod2 (green). ( C ) Quantification of distribution of GFP-positive cells in 5 even bins of cortex. ( D, E ) Quantification of GFP co- localization with Sox2+ ( D ) and Neurod2+ ( E ) cells. ( F ) Schematic model summarizing how loss of DDX3X-dependent translation impairs neurogenesis. Scale bars, indicated. Error bars, S.D. *p<0.05, ****p<0.0001.

Article Snippet: The following primary antibodies were used, rat: anti-SOX2 (Thermo Fisher, 14-9811-82, 1:1000), anti-BrdU (Abcam, ab6326, 1:200); mouse: anti-TUJ1 (Biolegend, 801202, 1:2000), anti-CTIP2 (Abcam, c8035, 1:500); rabbit: anti-TBR2 (Abcam, AB23345, 1:1000), anti-CC3 (Cell Signaling, 9661, 1:250), anti-NEUROD2 (Abcam, AB104430, 1:500), anti-Ki67 (Cell Signaling Technology, 12202, 1:1000), anti-PH3 (Millipore, 06-570, 1:500), anti-TBR1 (Cell Signaling Technology, 49661S, 1:1000), anti-Lhx2 (Millipore, ABE1402, 1:500), anti-DDX3X (Sigma Aldrich, HPA001648, 1:500); chicken anti-GFP (Abcam, Ab13970, 1:1000).

Techniques: Quantitative RT-PCR

Journal:

Article Title:

doi:

Figure Lengend Snippet: a Cumulative graph showing the log2 fold-change value of DEGs in dKAT3-ifKOs (mRNA-seq, 1 month after TMX, n = 3 per genotype). Upregulated genes are presented in red and downregulated genes in blue ( p .adj < 0.05 and |log2FC| ≥ 1). The bottom bar graph compares the area in each set. b Volcano plots of RNA-seq analysis. From left to right, we present all genes (left) and the subset of housekeeping genes listed in ref. (right). Gray: genes that are not significantly deregulated; red: upregulated genes; blue: downregulated genes; purple: housekeeping genes. c The 10 most enriched categories identified by Gene Ontology (GO) analysis on up- (red) and downregulated (blue) genes with a p .adj < 0.05 and |log2FC| ≥ 1. The upregulated gene set only retrieved three categories. d RNA-seq profiles for three representative neuronal-specific genes: Rbfox3 (NeuN), Hpca , and Camk4 . Scale: 2 kb. e Immunohistochemistry against the neuronal protein encoded by Camk4 , Rbfox3 (NeuN), and Hpca . Dashed line labels the position of the stratum pyramidale based on DAPI images ( n = 3). Scale: 60 μm. f Loss of NeuN immunoreactivity in DG neurons infected with a cre recombinase-expressing AAV (GFP + ). The experiment was performed 3 times with different sets of mice. The virus was injected unilaterally into the DG of adult Crebbp f/f ::Ep300 f/f mice and the mice were perfused 1 month later (Supplementary Fig. ). See Supplementary Fig. for immunostaining against CBP in the same brain slide. Scale: 200 μm. g Scheme representing the strategy to eliminate both KAT3 proteins in hippocampal PNCs from E17 dKAT3 f/f embryos. h Representative images showing morphological changes in Crebbp f/f ::Ep300 f/f hippocampal neurons infected with LV-CRE compared with LV-GFP control ( n = 6 in both groups). i RT-PCR demonstrates decreased levels of Neurod2 (ND2), Hpca (hippocalcin), and Rbfox3 (NeuN) transcripts in dKAT3-KO PNCs. In contrast, several housekeeping genes ( Ppl1 , Pgk1 , Rpl23 ) are unaffected ( n = 5–6 in both groups). Data are presented as mean values ± SEM. Two-tailed t -test: **** p < 0.0001, *** p < 0.001, ** p < 0.01. Source data for graphs in panel ( i ) are provided as a file.

Article Snippet: The following primary antibodies have been used in this study: anti-CBP, Santa Cruz sc-583 (IHC: 1:500; ChIP: 10 µg); anti-CBP, Santa Cruz sc-369 (ICC: 1:100); anti-CBP, Santa Cruz sc-7300 (IHC: 1:100; ICC: 1:100); anti-p300, Santa Cruz sc-585 (IHC: 1:100; ICC: 1:100; ChIP: 10 µg); anti-NeuroD2, Abcam ab109406 (ICC: 1:100); anti-H2Aac , (IHC: 1:100); anti-H2Bac , (IHC: 1:1000; ICC: 1:1000); anti-H3K9,14ac , (IHC: 1:400); anti-H3K27ac, Abcam ab4729 (IHC: 1:1000; ICC: 1:1000; ChIP: 5 µg); anti-H3K27me3 07-449 Millipore (IHC: 1:100); anti-H3K9me3 ab8898 Abcam (IHC: 1:100); anti-H4ac , (IHC: 1:100); anti-NeuN, MAB377 Millipore (IHC: 1:500; FANS: 1:500); anti-Hpca, Abcam ab24560 (IHC: 1:500; ICC: 1:500); anti-CaMKIV, BD Transduction Laboratories C28420 (IHC: 1:500); anti-Cleaved-Cas3, Cell Signalling #9661 (IHC: 1:200); anti-Fos, Synaptic Systems #226004 (IHC: 1:500); anti-mCherry/dsRed, Clontech 632496 (ICC: 1:1000); anti-GFP, Aves Labs GFP-1020 (IHC: 1:1000; ICC: 1:1000); anti-GFAP, Sigma G9269 (IHC: 1:200; ICC: 1:100); anti-H2A.Xγ, Abcam ab2893 (IHC: 1:200).

Techniques: RNA Sequencing Assay, Immunohistochemistry, Infection, Expressing, Injection, Immunostaining, Reverse Transcription Polymerase Chain Reaction, Two Tailed Test

Journal:

Article Title:

doi:

Figure Lengend Snippet: a Immunostaining against H3K27ac in the CA1 subfield of single and dKAT3 ifKOs (IF signal control: 21.62 ± 2.24, dKAT3-ifKOs: 4.94 ± 1.03, p < 0.0001, unpaired t -test); the single ifKOs show non-significant difference compared to controls. The experiment was repeated 3 times. Scale: 10 μm. b Genome-wide analysis of H3K27ac changes in dKAT3-ifKOs. Volcano plots show the fold change and significance values for all peaks (Global) and for the subsets of neuron-specific (Neuro) and pancellular peaks. c Classification of regulatory regions categorized by genomic features. d Metaplots of ATAC-seq, KAT3, and H3K27ac ChIP-seq signals in neuronal and pancellular enhancers in controls and dKAT3-ifKOs. Plots are centered in the peak center and expanded ±1 kb. e Correlation between length of the enhancer and expression level of the proximal gene. f Number of downregulated genes that contain enhancers or super-enhancers in control mice and percentage that lose acetylation in dKAT3-ifKOs. g Barplot showing the top 10 enriched categories from the Phenotype analysis for genes harboring neuronal enhancers. h TFBS analysis of neuronal (green), non-neuronal (red), and pancellular (orange) KAT3 peaks. Each motif family name is a user-curated approximation to the results provided by the MEME-suite algorithm. The most enriched de novo-identified binding motif in neuronal KAT3 (upper right motif) is very similar to the NeuroD2 motif (bottom right). Log E -values obtained with the expectation maximization algorithm (MEME-ChIP suite): Neuro: bHLH = 1130, GLI = 258, SOX = 257; Pancell: FOX = 130, ETS = 67, SP = 57; NoNeuro: ETV = 157, FOX = 105, SOX = 49. i TFBS analysis of neuronal regions with a reduced ATAC-seq signal in dKAT3-ifKOs. j RNA-seq profiles for Neurod2 and NeuroD6 . Scale: 1 kb. k Digital footprinting of NeuroD2 at neuronal and pancellular KAT3-bound regions. Values correspond to normalized Tn5 insertions. l Representative snapshots of KAT3, ATAC, and H3K27ac depletion with bHLH footprint and NeuroD2 overlaps, at two genes that are strongly downregulated in dKAT3-ifKOs. Source data for graphs in panels ( e ) and ( h ) are provided as a file.

Article Snippet: The following primary antibodies have been used in this study: anti-CBP, Santa Cruz sc-583 (IHC: 1:500; ChIP: 10 µg); anti-CBP, Santa Cruz sc-369 (ICC: 1:100); anti-CBP, Santa Cruz sc-7300 (IHC: 1:100; ICC: 1:100); anti-p300, Santa Cruz sc-585 (IHC: 1:100; ICC: 1:100; ChIP: 10 µg); anti-NeuroD2, Abcam ab109406 (ICC: 1:100); anti-H2Aac , (IHC: 1:100); anti-H2Bac , (IHC: 1:1000; ICC: 1:1000); anti-H3K9,14ac , (IHC: 1:400); anti-H3K27ac, Abcam ab4729 (IHC: 1:1000; ICC: 1:1000; ChIP: 5 µg); anti-H3K27me3 07-449 Millipore (IHC: 1:100); anti-H3K9me3 ab8898 Abcam (IHC: 1:100); anti-H4ac , (IHC: 1:100); anti-NeuN, MAB377 Millipore (IHC: 1:500; FANS: 1:500); anti-Hpca, Abcam ab24560 (IHC: 1:500; ICC: 1:500); anti-CaMKIV, BD Transduction Laboratories C28420 (IHC: 1:500); anti-Cleaved-Cas3, Cell Signalling #9661 (IHC: 1:200); anti-Fos, Synaptic Systems #226004 (IHC: 1:500); anti-mCherry/dsRed, Clontech 632496 (ICC: 1:1000); anti-GFP, Aves Labs GFP-1020 (IHC: 1:1000; ICC: 1:1000); anti-GFAP, Sigma G9269 (IHC: 1:200; ICC: 1:100); anti-H2A.Xγ, Abcam ab2893 (IHC: 1:200).

Techniques: Immunostaining, Genome Wide, ChIP-sequencing, Expressing, Binding Assay, RNA Sequencing Assay, Footprinting

Journal:

Article Title:

doi:

Figure Lengend Snippet: a Hippocampal PNCs from E17 Crebbp f/f :: Ep300 f/f embryos were co-transfected with constructs that drive the expression of the Cre recombinase and full-length CBP. b Plasmid combination to express recombinant CBP simultaneously to endogenous CBP and p300 ablation. c Representative images of NeuroD2 and hippocalcin staining in PNC transfected with the constructs shown in ( b ). Note the reduced expression in the GFP+ cells in the absence of heterologous full-length CBP (experiments in two independent PNCs). Scale: 10 μm. d Quantification of the percentage of NeuroD2-positive or -negative cells and hippocalcin-positive or -negative cells among all GFP-positive cells ( n = 60 neurons per condition). e Scheme of the CBP fragments used for rescuing NeuroD2 expression (see Methods for additional details). NLS: nuclear localization domain; KAT: acetyltransferase domain; KIX: kinase-inducible domain interacting domain; bHLH-i: region of interaction with bHLH transcription factors. f Hippocampal PNCs from E17 Crebbp f/f :: Ep300 f/f embryos were transfected with constructs that drive the expression of the Cre recombinase and the different CBP fragments and the KAT domain shown in panel ( e ). g Representative images of NeuroD2 staining in PNCs infected with LV-CREw/oGFP and transfected with the different domains of CBP shown in panel ( e ). PNCs infected with LV-GFP (i.e. with wild type phenotype) were used as a control for baseline NeuroD2 level (2 independent PNCs). Scale: 20 μm. h Quantification of the percentage of NeuroD2-positive or -negative cells among all GFP-positive neurons ( n = 30–80 neurons per condition). Source data for graphs in panels ( d ) and ( h ) are provided as a file.

Article Snippet: The following primary antibodies have been used in this study: anti-CBP, Santa Cruz sc-583 (IHC: 1:500; ChIP: 10 µg); anti-CBP, Santa Cruz sc-369 (ICC: 1:100); anti-CBP, Santa Cruz sc-7300 (IHC: 1:100; ICC: 1:100); anti-p300, Santa Cruz sc-585 (IHC: 1:100; ICC: 1:100; ChIP: 10 µg); anti-NeuroD2, Abcam ab109406 (ICC: 1:100); anti-H2Aac , (IHC: 1:100); anti-H2Bac , (IHC: 1:1000; ICC: 1:1000); anti-H3K9,14ac , (IHC: 1:400); anti-H3K27ac, Abcam ab4729 (IHC: 1:1000; ICC: 1:1000; ChIP: 5 µg); anti-H3K27me3 07-449 Millipore (IHC: 1:100); anti-H3K9me3 ab8898 Abcam (IHC: 1:100); anti-H4ac , (IHC: 1:100); anti-NeuN, MAB377 Millipore (IHC: 1:500; FANS: 1:500); anti-Hpca, Abcam ab24560 (IHC: 1:500; ICC: 1:500); anti-CaMKIV, BD Transduction Laboratories C28420 (IHC: 1:500); anti-Cleaved-Cas3, Cell Signalling #9661 (IHC: 1:200); anti-Fos, Synaptic Systems #226004 (IHC: 1:500); anti-mCherry/dsRed, Clontech 632496 (ICC: 1:1000); anti-GFP, Aves Labs GFP-1020 (IHC: 1:1000; ICC: 1:1000); anti-GFAP, Sigma G9269 (IHC: 1:200; ICC: 1:100); anti-H2A.Xγ, Abcam ab2893 (IHC: 1:200).

Techniques: Transfection, Construct, Expressing, Plasmid Preparation, Recombinant, Staining, Infection

Journal:

Article Title:

doi:

Figure Lengend Snippet: a Snap view of the Neurod2 locus. The profiles for CBP and p300 binding, H3K27 acetylation and ATAC-seq signal are shown. The bottom orange track corresponds to the NeuroD2 ChIP-seq data generated in ref. . A scheme presenting the strategy used to drive the KAT activity of p300 to the Neurod2 promoter using a fusion protein with dCas9 is also shown. The positions targeted by the gRNAs (red lines, gND2 A-D) are indicated. Note that the target regions are in the proximity of bHLH sites. b Scheme of the co-infection of LVs expressing cre recombinase, dCas9-KAT, and the Neurod2 gRNAs (A-D mix). c Representative image of NeuroD2 protein levels in the cells co-infected with LV-CRE-GFP and the lentiviruses LV-gRNA-mCherry specific for Neurod2 (white arrows). As a specificity control, we conducted the same experiment using a gRNA targeted to Hpca (blue arrows) ( n = 4 wells per condition in 2 PNCs). As a comparison, cells infected with LV-CRE-GFP alone (yellow arrows) show strongly diminished NeuroD2 levels as well. Scale bar: 50 μm. d Quantification of different cell subpopulations observed in the experiment shown in panel ( c ) ( n = 4 wells per condition in 2 PNCs). Two-tailed t -test: **** p < 0.0001, *** p < 0.001, ** p < 0.01; * p < 0.05. Data are presented as mean values ± SEM. e Scheme of rescue experiment with plasmids carrying dCas9-KAT and the Neurod2 gRNA in dKAT3-KO cells that had already lost Neurod2 expression. f Representative image of NeuroD2 expression in control (PNC + LV-GFP) and dKAT3-depleted (PNC + LV-CRE-GFP) neurons after transfection with dCas9-KAT and gND2-C targeting the NeuroD2 promoter. Arrows indicate LV-CRE-GFP infected cells transfected with the gRNA-carrying vector. A gRNA targeting the hippocalcin promoter was used as a specificity control. Scale: 20 μm. g Quantification of the percentage of transfected cells showing normal NeuroD2 expression after transfection with gND2-C alone and dCas9-KAT co-transfected with gND2-C, gND2-A, gND2-B, or gRNA control independently (experiments in three independent PNCs; n = 30–40 neurons per condition). Source data for graphs in panels ( d ) and ( g ) are provided as a file.

Article Snippet: The following primary antibodies have been used in this study: anti-CBP, Santa Cruz sc-583 (IHC: 1:500; ChIP: 10 µg); anti-CBP, Santa Cruz sc-369 (ICC: 1:100); anti-CBP, Santa Cruz sc-7300 (IHC: 1:100; ICC: 1:100); anti-p300, Santa Cruz sc-585 (IHC: 1:100; ICC: 1:100; ChIP: 10 µg); anti-NeuroD2, Abcam ab109406 (ICC: 1:100); anti-H2Aac , (IHC: 1:100); anti-H2Bac , (IHC: 1:1000; ICC: 1:1000); anti-H3K9,14ac , (IHC: 1:400); anti-H3K27ac, Abcam ab4729 (IHC: 1:1000; ICC: 1:1000; ChIP: 5 µg); anti-H3K27me3 07-449 Millipore (IHC: 1:100); anti-H3K9me3 ab8898 Abcam (IHC: 1:100); anti-H4ac , (IHC: 1:100); anti-NeuN, MAB377 Millipore (IHC: 1:500; FANS: 1:500); anti-Hpca, Abcam ab24560 (IHC: 1:500; ICC: 1:500); anti-CaMKIV, BD Transduction Laboratories C28420 (IHC: 1:500); anti-Cleaved-Cas3, Cell Signalling #9661 (IHC: 1:200); anti-Fos, Synaptic Systems #226004 (IHC: 1:500); anti-mCherry/dsRed, Clontech 632496 (ICC: 1:1000); anti-GFP, Aves Labs GFP-1020 (IHC: 1:1000; ICC: 1:1000); anti-GFAP, Sigma G9269 (IHC: 1:200; ICC: 1:100); anti-H2A.Xγ, Abcam ab2893 (IHC: 1:200).

Techniques: Binding Assay, ChIP-sequencing, Generated, Activity Assay, Infection, Expressing, Two Tailed Test, Transfection, Plasmid Preparation

Journal: Behavioural Neurology

Article Title: Effects of Extremely Low-Frequency Electromagnetic Fields on Neurogenesis and Cognitive Behavior in an Experimental Model of Hippocampal Injury

doi: 10.1155/2017/9194261

Figure Lengend Snippet: Densitometric analysis of NeuroD2 bands (at approximately 48 kDa) in hippocampal extracts of the study groups. (a and b) Representative graph of NeuroD2 expression in different groups ( ∗ P < 0.05).

Article Snippet: The membranes were blocked with 5% skim milk in TBS, containing 0.5% Tween 20, and were incubated with primary antibodies directed against mouse monoclonal anti-NeuroD2 antibody (1 : 1500; Abcam, Cambridge, UK) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 1 : 1000; Abcam, Cambridge, UK) for 2 h. After washing the membranes in a mixture of TBS and Tween 20 three times, they were reincubated with secondary alkaline-phosphatase-conjugated antibody (1 : 5000; Abcam, Cambridge, UK) for 1 h. Finally, the bands were detected with a chromogenic substrate (5-bromo-4-chloro-3-indolyl phosphate) in the presence of nitro blue tetrazolium.

Techniques: Expressing

Journal: BMC cardiovascular disorders

Article Title: Neurogenic differentiation 2 promotes inflammatory activation of macrophages in doxorubicin-induced myocarditis via regulating protein kinase D.

doi: 10.1186/s12872-025-04626-7

Figure Lengend Snippet: Fig. 4 Functional interaction between Neurogenic Differentiation 2 and protein kinase D. Macrophages were isolated from normal tissues and treated with 1µmol/L DOX for 12 h. (A) The binding site of PKD and Neurod2 was predicted by the JASPAR database. (B-C) qPCR analysis and western blotting were performed to detect the expression of Neurod2 in macrophages and tissues induced by DOX. (D-E) Macrophages isolated from the heart of normal mice were co-transfected with Neurod2 overexpression plasmid, and siNeurod2. qPCR and western blotting were performed to test the expression of Neurod2 mRNA and protein, respectively. (F-G) The levels of PKD in each group were explored by qPCR and western blotting. (H-I) The binding ability between PKD and Neurod2 was confirmed by dual-luciferase reporter assay (H) and CHIP assay (I). Values were presented as the mean ± SD of triplicate experiments. Compared with the control group or si-NC group, *P < 0.05, **P < 0. 01, ***P < 0. 001; Compared with the pc-NC group, #P < 0.05, ##P < 0. 01, ###P < 0. 001

Article Snippet: After blocking 1 h with 5% skimmed milk, the membranes were incubated with the following primary antibodies at 4°C overnight, including PKD (1:2000; Affinity Biosciences, Cincinnati, OH, USA), NLRP3 (1:2000; Affinity Biosciences), p-p65 (1:1000; Affinity Biosciences), p65 (1:1000; Affinity Biosciences), Neurod2 (1:1000; Biorbyt, Cambridge, UK.), GAPDH (1:3000; Affinity Biosciences).

Techniques: Functional Assay, Isolation, Binding Assay, Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation, Luciferase, Reporter Assay, Control

Journal: BMC cardiovascular disorders

Article Title: Neurogenic differentiation 2 promotes inflammatory activation of macrophages in doxorubicin-induced myocarditis via regulating protein kinase D.

doi: 10.1186/s12872-025-04626-7

Figure Lengend Snippet: Fig. 5 The overexpression of Protein kinase D can reverse the impact of neurogenic differentiation 2 on doxorubicin-induced inflammatory activation of macrophages. Macrophages isolated from normal tissue and treated with 1µmol/L DOX for 12 h. (A-B) Macrophages were co-transfected with PKD overexpression plasmid and si-Neurod2. qPCR was used to test the mRNA expressions of PKD and NLRP3. (C) The protein levels of PKD, p-p65, p65, and NLRP3 were detected. (D-G) The levels of TNF-α, IL-6, IL-18, and IL-1β in each group were measured by qPCR and ELISA. Values were presented as the mean ± SD of triplicate experiments. Compared with the DOX group, *P < 0.05, **P < 0. 01, ***P < 0. 001; Compared with DOX + pc-PKD group, #P < 0.05, ##P < 0. 01, ###P < 0. 001

Article Snippet: After blocking 1 h with 5% skimmed milk, the membranes were incubated with the following primary antibodies at 4°C overnight, including PKD (1:2000; Affinity Biosciences, Cincinnati, OH, USA), NLRP3 (1:2000; Affinity Biosciences), p-p65 (1:1000; Affinity Biosciences), p65 (1:1000; Affinity Biosciences), Neurod2 (1:1000; Biorbyt, Cambridge, UK.), GAPDH (1:3000; Affinity Biosciences).

Techniques: Over Expression, Activation Assay, Isolation, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

Journal: BMC cardiovascular disorders

Article Title: Neurogenic differentiation 2 promotes inflammatory activation of macrophages in doxorubicin-induced myocarditis via regulating protein kinase D.

doi: 10.1186/s12872-025-04626-7

Figure Lengend Snippet: Fig. 6 PKD-mediated macrophage inflammasome activation in heart tissue. The combination of NeuroD2 and PKD promoter could enhance NLRP3/ NF-κB signaling,

Article Snippet: After blocking 1 h with 5% skimmed milk, the membranes were incubated with the following primary antibodies at 4°C overnight, including PKD (1:2000; Affinity Biosciences, Cincinnati, OH, USA), NLRP3 (1:2000; Affinity Biosciences), p-p65 (1:1000; Affinity Biosciences), p65 (1:1000; Affinity Biosciences), Neurod2 (1:1000; Biorbyt, Cambridge, UK.), GAPDH (1:3000; Affinity Biosciences).

Techniques: Activation Assay

Journal: Immunity

Article Title: Prenatal interleukin 6 elevation increases glutamatergic synapse density and disrupts hippocampal connectivity in offspring

doi: 10.1016/j.immuni.2021.10.006

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were used: guinea pig anti-VGLUT1 (1:1500; Synaptic Systems); mouse anti-V-gat (Synaptic Systems, 1:1000), rabbit anti-IBA (WAKO, 1:200), mouse anti-GFAP (Sigma-Aldrich, 1:400), guinea pig anti-NeuN (Synaptic System, 1:500), mouse anti-SATB2 (AbCam,1:100), rabbit anti-NeuroD2 (AbCam, 1:1500).

Techniques: Recombinant, Software